CN1296356C - CCR-3 receptor antagonists VII - Google Patents

Abstract

The invention provides compounds of Formula (I): wherein: R1-R4, A, D, and L have any of the values defined in the specification that are CCR-3 receptor antagonists, pharmaceutical compositions containing them, methods for their use, and methods and intermediates useful for preparing them.

Description

CCR-3 receptor antagonist VII

The present invention relates to CCR-3 receptor antagonists, pharmaceutical compositions containing the antagonists, their use for treating diseases mediated by CCR-3, such as asthma, and a preparation method thereof.

Tissue eosinophilia is a feature of many pathological conditions such as asthma, rhinitis, eczema and parasitic infections (see Bousquet, J. et al., N. Eng. J. Med. 323:1033-1039 (1990) and Kay, A.B. and Corrigan, C.J., Br. Med. Bull. 48:51-64 (1992)). Accumulation and activation of eosinophils in asthma is associated with bronchial epithelial damage and hyperresponse to inotropic mediators. Chemokines such as RANTES, eotaxin and MCP-3 are known to activate eosinophils (see Baggiolini, M. and Dahinden, C.A., Immunol. Today. 15:127-133 (1994), Rot, A.M. et al., J.Exp. .Med.176, 1489-1495 (1992) and Ponath, P.D. et al., J.Clin.Invest., 97 volumes, #3, 604-612 (1996)).However, with RANTES that also induces migration of other types of leukocytes Unlike MCP-3, eotaxin is selectively chemotactic for eosinophils (see Griffith-Johnson, D.A. et al., Biochem. Biophy. Res. Commun. 197:1167 (1993) and Jose, P.J. et al., Biochem. Biophy .Res.Commun.207,788(1994)).No matter by subcutaneous or intraperitoneal injection or aerosol inhalation, the accumulation of specific eosinophils was observed at the site of eotaxin administration.(Referring to Griffith-Johnson, D.A. etc., Biochem.Biophy.Res.Commun.197: 1167 (1993); Jose, P.J. et al., J.Exp.Med.179, 881-887 (1994); Rothenberg, M.E. et al., J.Exp.Med.181, 1211( 1995) and Ponath, P.D., J. Clin. Invest., Vol. 97, #3, 604-612 (1996)).

Glucocorticoids such as dexamethasone, dehydrocortisone and hydrocortisone have been used to treat many eosinophil-associated diseases, including bronchial asthma (R.P. Schleimer et al., Am. Rev. Respir. Dis., 141, 559 (1990)). Glucocorticoids are thought to suppress IL-5, IL-3 mediated eosinophil survival in these diseases. However, prolonged use of glucocorticoids can lead to side effects such as glaucoma, osteoporosis and growth retardation in patients (see Hanania, N.A. et al., J.Allergy and Clin.Immunol., Vol. 96, 571-579 (1995) and Saha, M.T. et al., Acta Paediatrica, Vol. 86, #2, 138-142 (1997)). Therefore, there is a need for an alternative method of treating eosinophil-associated diseases without causing those undesired side effects.

Recently, the CCR-3 receptor was identified as the major chemokine receptor for eosinophils in their response to eotaxin, RANTES and MCP-3. When infected with a murine pre-beta lymphoma line, CCR-3 binds to eotaxin, RANTES and MCP-3 and renders these cells chemotactic responsive to eotaxin, RANTES and MCP-3 (see Ponath, P.D. et al., J. Exp. Med. 183, 2437-2448 (1996)). The CCR-3 receptor is expressed on the surface of eosinophils, T-cells (subtype Th-2), basophils and mast cells and is highly selective for eotaxin. Studies have shown that pretreatment of eosinophils with anti-CCR-3 mAb can completely inhibit the chemotaxis of eosinophils to eotaxin, RANTES and MCP-3 (see Heath, H. et al., J.Clin.Invest., 99 Vol. #2, 178-184 (1997)). Applicant's issued US Patents 6,140,344 and 6,166,015, as well as published EP application EP903349 (which was published on March 24, 1999), disclose CCR-3 antagonists that inhibit eosinophil recruitment by chemokines such as eotaxin.

Thus, blocking the ability of the CCR-3 receptor to bind RANTES, MCP-3, and eotaxin, thereby preventing the recruitment of eosinophils, would provide therapy for eosinophil-mediated inflammatory diseases.

The present invention relates to compounds capable of inhibiting the binding of eotaxin to CCR-3 receptors, thereby providing a method for resisting eosinophil-induced diseases such as asthma.

In a first aspect, the present invention provides compounds of formula (I):

in:

R ¹ is methylene or ethylene;

^R is optionally substituted phenyl;

^R is hydrogen, alkyl, acyl, aryl, or arylalkyl;

Ring A is cycloalkyl, heterocyclyl, or optionally substituted phenyl;

D is N or CR ^b ;

L is -C(=O)-, -C(=S)-, -SO ₂ -, -C(=O)N(R ^a )-, -C(=S)N(R ^a )-, - SO ₂ N(R ^a )-, -C(=O)O-, -C(=S)O-, -S(=O) ₂ O-;

R ^is alkyl, cycloalkyl, alkenyl, alkynyl, heteroalkyl or acylalkyl;

^R is hydrogen, alkyl, acyl, aryl, arylalkyl, alkoxycarbonyl, or benzyloxycarbonyl; and

R ^b is hydrogen or alkyl;

and prodrugs thereof, individual isomers, racemic and non-racemic mixtures of isomers, and pharmaceutically acceptable salts thereof.

Furthermore, among the compounds defined above (hereinafter they will be referred to as (i)), the following compounds are preferred:

(ii) a compound of (i), which is a compound of formula (II):

wherein R ¹ -R ⁴ , A, D and L are as defined in (i).

(iii) a compound of (i), which is a compound of formula (III):

wherein R ¹ -R ⁴ , A, D and L are as defined in (i).

(iv) The compound of any one of (i)-(iii), wherein R ¹ is methylene.

(v) The compound of any one of (i)-(iii), wherein R ² is 4-chlorophenyl or 3,4-dichlorophenyl.

(vi) The compound of any one of (i)-(iii), wherein R ³ is hydrogen.

(vii) The compound of any one of (i)-(iii), wherein -C(=O)-, -SO ₂ -, -C(=O)N(R ^a )-, -C(=S) N(R ^a )-, or -C(=O)O-.

(viii) a compound of (i), which is a compound of formula (IV):

wherein R ³ -R ⁴ , A, D and L are as defined in (i).

(ix) The compound of any one of (i)-(viii), wherein R ⁴ is cyclohexyl, allyl, isopropyl, n-butyl, or 2-(ethoxycarbonyl) ethyl.

(x) A compound of (i), wherein A is cyclopentyl.

(xi) A compound of (i) which is:

Cyclohexanecarboxylic acid {(1R,2R)-2-[4-(4-chloro-benzyl)-piperidin-1-yl]-cyclopentyl}-amide;

{(1R,2R)-2-[4-(4-Chloro-benzyl)-piperidin-1-yl]-cyclopentyl}-3-cyclohexyl-urea;

1-allyl-3-{(1R,2R)-2-[4-(4-chloro-benzyl)-piperidin-1-yl]-cyclopentyl}-urea;

1-{(1R,2R)-2-[4-(4-Chloro-benzyl)-piperidin-1-yl]-cyclopentyl}-3-isopropyl-urea;

1-Butyl-3-{(1R,2R)-2-[4-(4-chloro-benzyl)-piperidin-1-yl]-cyclopentyl}-urea;

3-(3-{(1R,2R)-2-[4-(4-Chloro-benzyl)-piperidin-1-yl]-cyclopentyl]-ureido)-propionic acid ethyl ester;

or its salt.

In a second aspect, the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

In a third aspect, the present invention provides a method for preparing the compound of formula (I) disclosed herein.

In a fourth aspect, the present invention provides novel intermediates disclosed herein for the preparation of compounds of formula (I).

In the fifth aspect, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in medical treatment or diagnosis, especially for treating diseases mediated by CCR-3, including respiratory diseases such as asthma.

In a sixth aspect, the present invention provides the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof for the preparation of a medicament for treating a disease (such as asthma) that can be treated by administering a CCR-3 receptor antagonist to a mammal .

Unless otherwise stated, the following terms used in this specification and claims have the following meanings:

"Acyl" means a group -C(O)R, where R is hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, phenyl or phenylalkyl, where alkyl, cycloalkyl, cycloalkyl Alkyl, and phenylalkyl are as defined herein. Typical examples include, but are not limited to, formyl, acetyl, cyclohexylcarbonyl, cyclohexylmethylcarbonyl, benzoyl, benzylcarbonyl.

"Acylalkyl" means an -alkylene-C(O)R group where R is hydrogen, alkyl, haloalkyl, cycloalkyl, cycloalkyl-alkyl, optionally substituted phenyl, benzyl radical, hydroxy, alkoxy, amino, monoalkylamino or dialkylamino. Typical examples include methylcarbonyl-methyl, 2-(ethoxycarbonyl)ethyl, 2-(methoxycarbonyl)ethyl, 2-carboxyethyl.

"Acylamino" means a -NR'C(O)R group, where R' is hydrogen or alkyl, and R is hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, phenyl, or phenylalkane wherein alkyl, cycloalkyl, cycloalkylalkyl, and phenylalkyl are as defined herein. Typical examples include, but are not limited to, formylamino, acetylamino, cyclohexylcarbonylamino, cyclohexylmethyl-carbonylamino, benzoylamino, benzylcarbonylamino.

"Alkoxy" means the group -OR where R is alkyl as defined herein, eg methoxy, ethoxy, propoxy, butoxy.

"Alkoxycarbonyl" means a group -C(O)-R where R is alkoxy as defined herein.

"Alkenyl" refers to a linear monovalent hydrocarbon group of 2-6 carbon atoms or a branched monovalent hydrocarbon group of 3-6 carbon atoms containing at least one double bond, such as vinyl, propenyl.

"Alkyl" refers to a straight-chain saturated monovalent hydrocarbon group of 1-6 carbon atoms or a branched saturated monovalent hydrocarbon group of 3-6 carbon atoms, such as methyl, ethyl, propyl, 2-propyl, n- Butyl, isobutyl, tert-butyl, pentyl.

"Alkylamino" or "monoalkylamino" refers to the radical -NHR, where R represents alkyl, cycloalkyl, or cycloalkyl-alkyl as defined herein. Typical examples include, but are not limited to, methylamino, ethylamino, isopropylamino, cyclohexylamino.

"Alkylene" refers to a straight-chain saturated divalent hydrocarbon group of 1-6 carbon atoms or a branched saturated divalent hydrocarbon group of 3-6 carbon atoms, such as methylene, ethylene, 2,2-dimethyl Ethylene, propylene, 2-methylpropylene, butylene, pentylene.

"Alkynyl" refers to a linear monovalent hydrocarbon group of 2-6 carbon atoms or a branched monovalent hydrocarbon group of 3-6 carbon atoms containing at least one triple bond, such as ethynyl, propynyl.

"Alkylsulfonyl" means a -S(O) ₂ R radical, where R is alkyl, cycloalkyl, or cycloalkylalkyl as defined herein, such as methylsulfonyl, ethylsulfonyl, propyl Sulfonyl, Butylsulfonyl, Cyclohexylsulfonyl.

"Alkylsulfinyl" means a -S(O)R radical, where R is alkyl, cycloalkyl, or cycloalkylalkyl as defined herein, such as methylsulfinyl, ethylsulfinyl, Propylsulfinyl, butylsulfinyl, cyclohexylsulfinyl.

"Alkylthio" means a -SR group where R is alkyl as defined above, eg methylthio, ethylthio, propylthio, butylthio.

"Aryl" refers to a monocyclic or bicyclic aromatic hydrocarbon group of 6-10 ring atoms, which is optionally substituted by one or more substituents, preferably one, two or three substituents, which are preferably selected from Alkyl, haloalkyl, hydroxyalkyl, heteroalkyl, acyl, amido, amino, alkylamino, dialkylamino, alkylthio, alkylsulfinyl, alkylsulfonyl, -SO ₂ NR'R" (where R' and R" are independently hydrogen or alkyl), alkoxy, haloalkoxy, alkoxycarbonyl, carbamoyl, hydroxyl, halogen, nitro, cyano, mercapto, methylenedioxy base or ethylenedioxy. More specifically, the term aryl includes, but is not limited to, phenyl, chlorophenyl, fluorophenyl, methoxyphenyl, 1-naphthyl, 2-naphthyl, and derivatives thereof.

"Arylene" refers to a divalent aryl group as defined above.

"Aralkyl" means an alkyl group as defined herein in which one hydrogen atom of the alkyl group is replaced by an aryl group. Typical aralkyl groups include, but are not limited to, benzyl, 2-phenyleth-1-yl, naphthylmethyl, 2-naphthyleth-1-yl, naphthylbenzyl, 2-naphthylpheneth-1-yl .

"Aryloxy" means the group -O-R, where R is aryl as defined herein.

"Carbamoyl" refers to a -C(=O) _NH2 group.

"Cycloalkyl" refers to a saturated monovalent cyclic hydrocarbon group of 3-7 ring carbons, such as cyclopropyl, cyclobutyl, cyclohexyl, 4-methylcyclohexyl.

"Cycloalkylalkyl" means a group -R ^x R ^y where R ^x is alkylene and R ^y is cycloalkyl as defined herein, eg cyclohexylmethyl.

"Dialkylamino" means the group -NRR', wherein R and R' independently represent alkyl, cycloalkyl, or cycloalkylalkyl as defined herein. Typical examples include, but are not limited to, dimethylamino, methylethylamino, bis(1-methylethyl)amino, (cyclohexyl)(methyl)amino, (cyclohexyl)(ethyl)amino, (cyclohexyl )(propyl)amino, (cyclohexylmethyl)(methyl)amino, (cyclohexylmethyl)(ethyl)amino.

"Halogen" means fluorine, chlorine, bromine, or iodine, preferably fluorine and chlorine.

"Haloalkyl" refers to an alkyl group substituted with one or more same or different halogen atoms, such as -CH ₂ Cl, -CF ₃ , -CH ₂ CF ₃ , -CH ₂ CCl ₃ .

"Heteroaryl" refers to a monocyclic or bicyclic group containing at least one aromatic ring of 5-12 ring atoms, which contains one, two, or three ring heteroatoms selected from N, O, or S, The remaining ring atoms are C with the proviso that the point of attachment of the heteroaryl group will be on the aromatic ring. The heteroaromatic ring is optionally substituted independently by one or more substituents, preferably one or two substituents, selected from the group consisting of alkyl, haloalkyl, hydroxyalkyl, heteroalkyl, acyl, amido, Amino, alkylamino, dialkylamino, alkylthio, alkylsulfinyl, alkylsulfonyl, -SO ₂ NR'R" (where R' and R" are independently hydrogen or alkyl), alkyl Oxy, haloalkoxy, alkoxycarbonyl, carbamoyl, hydroxy, halogen, nitro, cyano, mercapto, methylenedioxy or ethylenedioxy. More specifically, the term heteroaryl includes, but is not limited to, pyridyl, furyl, thienyl, thiazolyl, isothiazolyl, triazolyl, imidazolyl, isoxazolyl, pyrrolyl, pyrazolyl, pyrimidinyl, Benzofuryl, tetrahydrobenzofuryl, isobenzofuryl, benzothiazolyl, benzisothiazolyl, benzotriazolyl, indolyl, isoza(hetero)indenyl, benzoxa Azolyl, quinolinyl, tetrahydroquinolyl, isoquinolyl, benzimidazolyl, benzisoxazolyl or benzothienyl and derivatives thereof.

"Heteroarylene" refers to a divalent heteroaryl group as defined above.

"Heteroaralkyl" means an alkyl group as defined herein, wherein one hydrogen atom of the alkyl group is replaced by a heteroaryl group as defined herein.

"Heteroalkyl" means an alkyl group as defined herein, wherein one, two or three hydrogen atoms are independently selected from -OR ^a , -NR ^b R ^c , and -S(O) _n R ^d (where n is an integer from 0 to 2), with the proviso that the point of attachment of the heteroalkyl is through a carbon atom of the heteroalkyl. R ^a is hydrogen, acyl, alkyl, cycloalkyl or cycloalkylalkyl. R ^b and R ^c are independently hydrogen, acyl, alkyl, cycloalkyl or cycloalkylalkyl; when n is 0, R ^d is hydrogen, alkyl, cycloalkyl, cycloalkylalkyl; When n is 1 or 2, ^R is alkyl, cycloalkyl, cycloalkylalkyl, amino, amido, monoalkylamino or dialkylamino; typical examples include, for example, 2-hydroxyethyl, 3-hydroxypropyl, 2-hydroxy-1-hydroxymethylethyl, 2,3-dihydroxypropyl, 1-hydroxymethylethyl, 3-hydroxybutyl, 2,3-dihydroxybutyl, 2-Hydroxy-1-methylpropyl, 2-aminoethyl, 3-aminopropyl, 2-methylsulfonylethyl, aminosulfonylmethyl, aminosulfonylethyl, aminosulfonylpropyl, Methylaminosulfonylmethyl, methylaminosulfonylethyl, methylaminosulfonylpropyl.

"Heterocyclyl" means a saturated or unsaturated non-aromatic ring group having 3-8 ring atoms, one or two of which are selected from NR ^x {wherein each R ^x is independently hydrogen, alkyl, acyl , alkylsulfonyl, aminosulfonyl, (alkylamino)sulfonyl, (dialkylamino)sulfonyl, carbamoyl, (alkylamino)carbonyl, (dialkylamino)carbonyl, (carbamoyl )alkyl, (alkylamino)carbonylalkyl, or dialkylaminocarbonylalkyl}, O, or a heteroatom of S(O) _n (where n is an integer from 0 to 2), and the remaining ring atoms are carbon . The heterocyclyl ring may be optionally substituted independently with one, two or three substituents selected from the group consisting of alkyl, haloalkyl, heteroalkyl, halo, nitro, cyanoalkyl, hydroxy, alkoxy , amino, monoalkylamino, dialkylamino, aralkyl, -(X) _n -C(O)R (wherein X is O or NR', n is 0 or 1, R is hydrogen, alkyl, Haloalkyl, hydroxy, alkoxy, amino, monoalkylamino, dialkylamino or optionally substituted phenyl, and R' is hydrogen or alkyl), -alkylene-C(O)R (where R is hydrogen, alkyl, haloalkyl, hydroxy ^{, alkoxy, amino, monoalkylamino, dialkylamino or optionally substituted phenyl) or -S(O)nRd} ₍ wherein n is 0- 2, ^Rd is hydrogen (provided that n is 0), alkyl, haloalkyl, cycloalkyl, cycloalkylalkyl, amino, monoalkylamino, dialkylamino, or hydroxyalkyl). More specifically, the term heterocyclyl includes, but is not limited to, tetrahydropyranyl, piperidino, N-methylpiperidin-3-yl, piperazino, N-methylpyrrolidin-3-yl, 3-pyrrolidino, morpholino, thiomorpholino, thiomorpholino-1-oxide, thiomorpholino-1,1-dioxide, tetrahydrothiophene-S, S- Dioxide, pyrrolinyl, imidazolinyl, and their derivatives.

"Hydroxyalkyl" means an alkyl group as defined herein substituted with one or more, preferably one, two or three hydroxy groups, provided that the same carbon atom does not carry more than one hydroxy group. Typical examples include, but are not limited to, 2-hydroxyethyl, 2-hydroxypropyl, 3-hydroxypropyl, 1-(hydroxymethyl)-2-methylpropyl, 2-hydroxybutyl, 3-hydroxybutyl base, 4-hydroxybutyl, 2,3-dihydroxypropyl, 2-hydroxy-1-hydroxymethylethyl, 2,3-dihydroxybutyl, 3,4-dihydroxybutyl and 2-( Hydroxymethyl)-3-hydroxypropyl, preferably 2-hydroxyethyl, 2,3-dihydroxypropyl and 1-(hydroxymethyl)-2-hydroxyethyl. Accordingly, the term "hydroxyalkyl" as used herein is used to define a subset of heteroalkyl groups.

"Leaving group" has the meaning conventionally associated with synthetic organic chemistry, i.e. an atom or group capable of being displaced by a nucleophile, including halogens (such as chlorine, bromine, and iodine), alkanesulfonyloxy, arylsulfonyl Oxygen, alkylcarbonyloxy (such as acetyloxy), arylcarbonyloxy, methanesulfonyloxy, tosyloxy, trifluoromethanesulfonyloxy, aryloxy (such as 2,4- dinitrophenoxy), methoxy, N,O-dimethylhydroxylamino.

"Optionally substituted phenyl" means phenyl optionally substituted with one or more substituents, preferably one, two or three substituents, preferably selected from the group consisting of alkyl, haloalkyl, hydroxyalkyl, Heteroalkyl, acyl, acylamino, amino, alkylamino, dialkylamino, alkylthio, alkylsulfinyl, alkylsulfonyl, -SO ₂ NR'R" (where R' and R" are independently hydrogen or alkyl), alkoxy, haloalkoxy, alkoxycarbonyl, carbamoyl, hydroxy, halogen, nitro, cyano, mercapto, methylenedioxy or ethylenedioxy. More specifically, the term includes, but is not limited to, phenyl, chlorophenyl, fluorophenyl, bromophenyl, methylphenyl, ethylphenyl, methoxyphenyl, cyanophenyl, 4-nitrophenyl phenyl, 4-trifluoromethylphenyl, 4-chlorophenyl, 3,4-difluorophenyl, 2,3-dichlorophenyl, 3-methyl-4-nitrophenyl, 3 -Chloro-4-methylphenyl, 3-chloro-4-fluorophenyl or 3,4-dichlorophenyl and derivatives thereof.

"Optional" or "optionally" means that the subsequently described event or circumstance can but need not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not. For example, "aryl optionally mono- or di-substituted with alkyl" means that the alkyl group may but need not be present, and the description includes cases where the aryl group is mono- or di-substituted with an alkyl group, and aryl The case where the group is not substituted by an alkyl group.

"Pharmaceutical excipient" refers to an excipient used in the preparation of a pharmaceutical composition, which is generally safe, non-toxic and has neither biological nor other adverse reactions, including excipients The formulation is qualified for both veterinary use and human pharmaceutical use. A "pharmaceutically acceptable excipient" as used in the specification and claims includes both one and more than one such excipient.

A "pharmaceutically acceptable salt" of a compound refers to a salt which is pharmaceutically acceptable and which possesses the desired pharmaceutical activity of the parent compound. Such salts include: (1) acid addition salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc.; Acetic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3-(4-hydroxybenzoyl)-benzene Formic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid acid, 4-toluenesulfonic acid, camphorsulfonic acid, 4-methylbicyclo[2.2.2]oct-2-ene-1-carboxylic acid, glucoheptonic acid, 3-phenylpropionic acid, trimethylacetic acid, Acid addition salts formed by tert-butylacetic acid, lauryl sulfate, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, muconic acid; or (2) when the acidic protons present in the parent compound Substituted by metal ions, such as alkali metal ions, alkaline earth metal ions, or aluminum ions; or formed by coordination with organic bases such as ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methylglucamine of salt.

"Phenylalkyl" means an alkyl group as defined herein, wherein one hydrogen atom of the alkyl group is replaced by an optionally substituted phenyl group.

"Protecting group" refers to a grouping of atoms that, when attached to a reactive group in a molecule, masks, reduces or prevents its reactivity. Examples of protecting groups can be found in TWGreen and PGFuts, Protective Groups in Organic Chemistry , (Wiley, Second Edition, 1991) and Harrison and Harrison et al., Compendium of Synthetic Organic Methods , Vol. 1-8 (John Wiley & Sons, 1971-1996) found in. Typical amino protecting groups include formyl, acetyl, trifluoroacetyl, benzyl, benzyloxycarbonyl (CBZ), tert-butoxycarbonyl (Boc), trimethylsilyl (TMS), 2-trimethylsilyl Amyl-ethanesulfonyl (SES), trityl and substituted trityl, allyloxycarbonyl, 9-fluorenylmethoxycarbonyl (FMOC), nitro-veratroxycarbonyl (NVOC). Typical hydroxyl protecting groups include those in which the hydroxyl group is acylated or alkylated, such as benzyl, and trityl ethers as well as alkyl ethers, tetrahydropyranyl ethers, trimethylsilyl ethers, and allyl ethers. ether.

"Treatment" or "therapy" of disease includes: (1) prophylaxis of disease, i.e. causing clinical symptoms of disease not to develop in mammals that may be exposed to or susceptible to disease but have not yet experienced or shown symptoms of disease (2) inhibiting the disease, that is, preventing or reducing the development of the disease or its clinical symptoms; or (3) alleviating the disease, that is, promoting the regression of the disease or its clinical symptoms.

A "therapeutically effective amount" refers to an amount of a compound sufficient to effect treatment of a disease when administered to a mammal to treat the disease. The "therapeutically effective amount" will vary depending on the compound, the disease and its severity and the age, weight, etc. of the mammal to be treated.

"Prodrugs" refer to compounds that release the active parent drug according to formula (I) in vivo when these prodrugs are administered to a mammalian subject. Prodrugs of compounds of formula (I) are prepared by modifying functional groups present on compounds of formula (I) in such a way that the modification is cleaved in vivo, releasing the parent compound. Prodrugs include compounds of formula (I) in which the hydroxy, amino or thiol groups of the compound of formula (I) are bound to any group that cleaves in vivo to regenerate a free hydroxy, amino or sulfhydryl group, respectively. Examples of prodrugs include, but are not limited to, esters (e.g. acetate, formate, benzoate derivatives) of hydroxyl functional groups in compounds of formula (I), carbamates (e.g. N,N-dimethyl aminocarbonyl) etc.

Compounds that have the same molecular formula but differ in the nature or sequence of bonding of their atoms, or the arrangement of their atoms in space, are termed "isomers". Isomers that differ in the arrangement of their atoms in space are termed "stereoisomers". Stereoisomers that are not mirror images of each other are termed "diastereomers", and stereoisomers that are non-superimposable mirror images of each other are termed "enantiomers". When a compound has an asymmetric center, such as a carbon atom connecting four different groups, there can be a pair of enantiomers. Enantiomers can be characterized by the absolute configuration of their asymmetric centers, by the R- and S-sequence rules of Cahn and Prelog, or by the way in which the molecule rotates the plane of polarized light and is designated as dextrorotatory or levorotatory (i.e. are described as (+) or (-)-isomers, respectively). A chiral compound can exist as individual enantiomers or as a mixture thereof. A mixture containing equal proportions of the enantiomers is termed a "racemic mixture".

The compounds of the present invention may possess one or more asymmetric centers; such compounds may thus be produced as individual (R)- or (S)-stereoisomers or as mixtures thereof. Unless otherwise stated, throughout the specification and claims, the description or designation of a particular compound is meant to include both the individual enantiomers and their racemic or other mixtures. Methods for determination of stereochemistry and separation of stereoisomers are well known in the art (see discussion in Chapter 4 of "Advanced Organic Chemistry", 4th Edition J. March, John Wiley and Sons, New York, 1992) .

In general, the nomenclature used in this application is based on AUTONOM( ^TM) , a computer system of the Beilstein Institute for generating IUPAC systematic nomenclature. For example, R ₁ is methylene; R ₂ is 4-chlorophenyl; L is C(=O); A is cyclopentyl; R ₃ is hydrogen; R ⁴ is cyclohexyl; and D is -CH-( The compound of formula (I) of Example 1), named cyclohexanecarboxylic acid {2-[4-(4-chloro-benzyl)-piperidin-1-yl]-cyclopentyl}-amide .

Typical compounds of formula (I) are shown in the table below.

While the broadest definition of the invention is described above, certain compounds of formula (I) are preferred.

A preferred compound of the invention is a compound of formula (I) wherein R ₁ is methylene.

Another preferred compound of the invention is a compound of formula (I) wherein ring A is cyclopentyl. It was unexpectedly found that compounds in which ring A is cyclopentyl bind strongly to the CCR-3 receptor. Another preferred compound of the present invention is that wherein ring A is heterocyclyl (especially tetrahydropyranyl, S, S-dioxo-tetrahydrothiophenyl, tetrahydrothiophenyl (thiophenyl) or pyrrolidinyl) A compound of formula (I) or a compound of formula (I) wherein ring A is phenyl.

A preferred compound of the present invention is a compound of formula (I) wherein R is a benzene ring substituted by ^one , or two, substituents selected from the group consisting of alkyl, alkoxy, haloalkyl, halogen, cyano radical or nitro; preferably methyl, ethyl, methoxy, trifluoromethyl, chlorine, fluorine or bromine; most preferably 4-nitrophenyl, 4-trifluoromethylphenyl, 4-chlorophenyl , 3,4-difluorophenyl, 2,3-dichlorophenyl, 3-methyl-4-nitrophenyl, 3-chloro-4-methylphenyl, 3-chloro-4-fluorobenzene or 3,4-dichlorophenyl; particularly preferably 4-chlorophenyl or 3,4-dichlorophenyl.

A preferred compound of the invention is a compound of formula (I) wherein ^R3 is hydrogen or methyl, preferably hydrogen.

A preferred compound of the invention is that wherein L is -C(=O)-, -SO ₂ -, -C(=O)N(R ^a )-, -C(=S)N(R ^a )- or Compounds of formula (I) -C(=O)O-. More preferred are compounds wherein L is -C(=O)-, -C(=O)N(R ^a )-, most preferred are those wherein L is -C(=O)N(R ^a )- compound. Wherein R ^a is preferably hydrogen or methyl, most preferably hydrogen.

A preferred compound of the invention is a compound of formula (I) wherein D is N. When D is ^CRb , preferred compounds are those wherein ^Rb is hydrogen.

A preferred compound of the present invention is wherein R is ^alkyl , cycloalkyl, cycloalkylalkenyl or acylalkyl; more preferably cyclohexyl, allyl, isopropyl, n-butyl, or 2- (Ethoxycarbonyl)ethyl compounds of formula (I).

A specific compound of formula (I) is a compound of formula (II):

wherein R ¹ -R ⁴ , A, D, and L have any of the definitions described herein.

A specific compound of formula (I) is the compound of formula (III):

wherein R ¹ -R ⁴ , A, D, and L have any of the definitions described herein.

A specific compound of formula (I) is a compound of formula (IV):

Wherein R ³ , R ⁴ , A, D, and L have any definitions described in the present invention.

A specific compound of formula (I) is a compound of formula (V):

Wherein R ⁴ and D have any definition described in the present invention.

A specific compound of formula (I) is a compound of formula (VI):

Wherein R ⁴ and D have any definition described in the present invention.

A specific compound of formula (I) is the compound of formula (VII):

Wherein R ⁴ and D have any definition described in the present invention.

A specific compound of formula (I) is the compound of formula (VIII):

wherein R ^and D have any of the definitions described herein; ^R is hydrogen, alkyl, acyl, alkylsulfonyl, aminosulfonyl, (alkylamino)sulfonyl, (dialkylamino)sulfonyl, Carbamoyl, (alkylamino)carbonyl, (dialkylamino)carbonyl, (carbamoyl)alkyl, (alkylamino)carbonylalkyl, or dialkylaminocarbonylalkyl.

A specific compound of formula (I) is a compound of formula (IX):

Wherein R ⁴ and D have any definition described in the present invention.

A specific compound of formula (I) is a compound of formula (X):

Wherein R ⁴ and D have any definition described in the present invention.

Particularly preferred compounds of the invention are:

Cyclohexanecarboxylic acid {(1R,2R)-2-[4-(4-chloro-benzyl)-piperidin-1-yl]-cyclopentyl}-amide;

{(1R,2R)-2-[4-(4-Chloro-benzyl)-piperidin-1-yl]-cyclopentyl}-3-cyclohexyl-urea;

1-allyl-3-{(1R,2R)-2-[4-(4-chloro-benzyl)-piperidin-1-yl]-cyclopentyl}-urea;

1-{(1R,2R)-2-[4-(4-Chloro-benzyl)-piperidin-1-yl]-cyclopentyl}-3-isopropyl-urea;

1-Butyl-3-{(1R,2R)-2-[4-(4-chloro-benzyl)-piperidin-1-yl]-cyclopentyl}-urea;

3-(3-{(1R,2R)-2-[4-(4-Chloro-benzyl)-piperidin-1-yl]-cyclopentyl}-ureido)-propionic acid ethyl ester;

or its salt.

The compound of the present invention is a CCR-3 receptor antagonist, and inhibits eosinophil recruitment induced by CCR-3 chemokines such as RANTES, eotaxin, MCP-2, MCP-3 and MCP-4. The compounds of the present invention and compositions containing them are useful in the treatment of eosinophil-induced diseases, such as inflammatory or allergic diseases, and include respiratory allergic diseases such as asthma, allergic rhinitis, allergic lung disease, Hypersensitivity pneumonitis, eosinophilic pneumonitis (eg, chronic eosinophilic pneumonia); inflammatory bowel disease (eg, Crohn's disease and ulcerative colitis); and psoriasis and inflammatory skin disorders such as dermatitis and eczema .

The CCR-3 antagonistic activity of the compounds of the present invention can be determined by in vitro assays, such as ligand binding assays and chemotaxis assays, which are described in more detail in Examples 4, 5 and 6. In vivo testing can be performed by ovalbumin-induced asthma in the Balb/c mouse model, which is described in more detail in Example 7.

In general, the compounds of the present invention are administered in a therapeutically effective amount according to any recognized route of administration for pharmaceutical agents suitable for the same application. The actual amount of the compound, ie, active ingredient, of the invention will depend on such factors as the severity of the condition being treated, the age and relative health of the subject, the potency of the compound employed, the route and form of administration, and other factors.

A therapeutically effective amount of a compound of formula (I) may be about 0.01-20 mg per kilogram body weight of the recipient per day; preferably about 0.1-10 mg/kg/day. Thus, for administration to a 70 kg human, the most preferred dosage range is about 7 mg to 0.7 g per day.

Typically, the compounds of the invention are administered as pharmaceutical compositions by any of the following routes: oral, transdermal, inhalation (eg intranasal or oral inhalation) or parenteral (eg intramuscular, intravenous or subcutaneous). The preferred way of administration is oral administration, which can be adjusted according to the severity of the disease. The compositions can be formulated as tablets, pills, capsules, semi-solids, powders, sustained release formulations, solutions, suspensions, liposomes, elixirs, or any other suitable composition. Another preferred mode for administering the compounds of the invention is inhalation. This is an effective means for delivering therapeutic agents directly to the respiratory tract to treat diseases such as asthma and other similar or related respiratory diseases (see US Patent No. 5,607,915).

The choice of dosage form is based on factors such as the mode of administration of the drug and the bioavailability of the drug. For delivery by inhalation, the compounds can be formulated as liquid solutions or suspensions, aerosol propellants or dry powders filled with suitable dispersants for oral administration. There are three types of pharmaceutical inhalation devices: nebulizer inhalers, metered dose inhalers (MDI), and dry powder inhalers (DPI). A nebulizer device produces a high velocity air stream which sprays a therapeutic drug (which has been formulated into a liquid dosage form) into a spray which enters the patient's airways. MDIs typically package the formulation with compressed gas. Upon stimulation, the device releases a dose of the therapeutic drug via compressed gas, thereby providing a reliable method of administering a dose of the drug. DPI dosage form is a free-flowing powder dosage form of the drug that can be dispersed into the patient's respiratory airflow when the device is inhaled. The therapeutic agent can be formulated with excipients such as lactose in order to obtain a free-flowing powder. Measurable amounts of drug are stored in a capsule dosage form and dispensed to the patient with each activation. Recently, pharmaceutical formulations have been developed especially for drugs with poor bioavailability, based on the principle that bioavailability can be increased by increasing surface area, ie decreasing particle size. For example, US Patent No. 4,107,288 describes pharmaceutical formulations in the particle size range of 10-1,000 nm in which the active substance is loaded onto a polymeric cross-linked backbone. U.S. Patent No. 5,145,684 describes a pharmaceutical formulation in which the drug is ground into nanoparticles (average particle size 400 nm) in the presence of surface modifiers and then dispersed in a liquid medium to obtain pharmaceutical dosage forms.

Typically, the above compositions comprise a compound of formula (I) in combination with at least one pharmaceutically acceptable excipient. Acceptable excipients are non-toxic, facilitate administration and do not adversely affect the therapeutic benefit of the compound of formula (I). The excipient may be any solid, liquid, semi-solid or gaseous excipient in aerosol compositions generally available to those skilled in the art.

Solid pharmaceutical excipients include starch, cellulose, talc, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, magnesium stearate, sodium stearate, glycerol monostearate, Sodium chloride, dry skim milk, etc. Liquid and semisolid excipients can be selected from glycerol, propylene glycol, water, and various oils, including those of petroleum, animal, vegetable, or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil, and the like. Preferred liquid carriers, especially for injection solutions, include water, saline, glucose solution, glycols and the like.

Compressed gases can be used to disperse the compounds of the invention in aerosol dosage forms. Inert gases suitable for this purpose are nitrogen, carbon dioxide and the like.

For liposomal dosage forms for parenteral or oral drug delivery, the lipid is dissolved in a suitable organic solvent, eg tert-butanol, cyclohexane (1% ethanol). The solution is lyophilized and the lipid mixture is dispersed into a buffer solution that allows liposome formation. If necessary, liposomes can be reduced in size by sonication. (See Frank Szoka, Jr. and Demetrios Papahadjopoulos, "Comparative properties and methods of lipid vesicle (liposome) formulations", Ann.Rev.Biophys.Bioeng., 9:467-508 (1980), and D.D.Lasic" New Applications of Liposomes" Trends in Biotech., 16:467-608, (1998)).

Other suitable pharmaceutical excipients and their dosage forms are described in Remington's Pharmaceutical Sciences, edited by E.W. Martin (Mack Publishing Company, 18th Edition, 1990).

The amount of the compound in the formulation may vary within the range normally used by those skilled in the art. Usually, the preparation contains 0.01-99.99% by weight of the compound of formula (I) (based on the total weight of the preparation), and the rest is supplemented by one or more suitable pharmaceutical excipients. Preferably, the compound is present in an amount of 1-80 wt%. A representative formulation containing a compound of formula (I) is described in Example 4.

Those skilled in the art understand that the compounds of the present invention can be prepared in a number of ways. Preferred methods include, but are not limited to, the general synthetic procedures described below.

The starting materials and reagents used to prepare these compounds are available from commercial suppliers such as Aldrich Chemical Company (Milwaukee, WI, USA), Bachem (Torrance, CA, USA), Enika-Chemie, or Sigma (USA St. Louis, Missouri), Maybridge (Dist: Ryan Scientific, P.O. Box 6496, Columbia, S.C. 92960), Bionet Research Ltd., (Cornwall PL32 9QZ, UK), Menai Organics Ltd., (Gwynedd, N.Wales, UK), Butt Park Ltd., (Dist. Interchim, Montlucon Cedex, France), or prepared by methods known to those skilled in the art following the steps set forth in the literature, such as Fieser and Fieser' reagents for organic synthesis, Volumes 1-17 (John Wiley and Sons, 1991); Rodd's Chemistry of Carbon Compounds, Volumes 1-5 and Appendix (Elsevier Science Publishers, 1989), Organic Reactions, Volumes 1-40 (John Wiley and Sons, 1991), March's Advanced Organic Chemistry, (John Wiley and Sons, 1992), and Larock's Encyclopedia of Organic Transformations (VCH Publishers Inc., 1989). These scheme schemes merely illustrate some of the ways in which the compounds of the invention may be synthesized and modifications or suggestions to these schemes may be made by those skilled in the art having regard to this disclosure.

Starting materials and reaction intermediates can be isolated and purified, if necessary, using conventional techniques including, but not limited to, filtration, distillation, crystallization, chromatography, and the like. These materials can be characterized using conventional methods, including physical constants and spectral data.

Typically, compounds of formula (I) are prepared from precursor amines of formula (Ia):

The preparation of compounds of formula (Ia) and their conversion to compounds of formula (I) are described in Schemes 1-8 below.

Schemes 1-5 show the preparation of compounds of formula Ia with different ring A's. Provided in Preparations 1-6 are specific examples where R ¹ -R ² are 4-chlorobenzyl. The preparation of analogous compounds wherein ^R1 and ^R2 are varied within the scope of the invention is readily prepared by a person skilled in the art following this specification and references therein.

Reaction Scheme 1. Synthesis of Cyclobutylamine - Ring A = Cyclobutyl

R ₂ N=4-(4-chlorobenzyl)piperidine or 1-(4-chlorobenzyl)piperazine

Reaction scheme 2. Synthesis of cis-diamine-ring A=cyclopentyl and cyclohexyl.

R ₂ N=4-(4-chlorobenzyl)piperidine or 1-(4-chlorobenzyl)piperazine

Reaction scheme 3. Synthesis of transdiamine - ring A = cycloalkyl, tetrahydrofuranyl, pyrrolidinyl or tetrahydrothiophenyl

General Method A: (Alkylation of Amines with Epoxides)

0.5-1.5M amine solution, R ₂ NH (1 eq), and the designated epoxide, 3a (1.1-10 eq) were placed in EtOH, stirred at 80-95°C for 2-4.5 days, cooled to room temperature, concentrate. Crude aminoalcohols are purified by chromatography or recrystallization.

General Procedure B: (Amine Formation Using Methanesulfonyl Chloride and Ammonium Hydroxide)

A 0.2-0.3 M solution of the aminoalcohol (1 eq) in _CH2Cl2 at ₀ °C was treated successively with _Et3N (2 eq) and _MeSO2Cl (2 eq) and stirred at 0 °C for 1-2 hours, partitioned between _CH2Cl2 and _10-15 % _NH4OH . The aqueous phase was extracted with _CH2Cl2 , _the extract was dried and concentrated. A 0.13M solution of the residue in 2.5:1 dioxane:28-30 wt% _NH4OH was stirred at 70-80°C for 2.5-18 hours, cooled to room temperature, and concentrated. The residue was partitioned between EtOAc and 1N NaOH, the aqueous phase was extracted with EtOAc, the extract was washed with brine, dried and concentrated. The crude product was purified by chromatography or used without further purification.

Reaction Scheme 4. Synthesis of Sulfone - Ring A = Sulfolane (Is it correct??)

R ₂ N=4-(4-chlorobenzyl)piperidine or 1-(4-chlorobenzyl)piperazine

Reaction Scheme 5. Synthesis of Aniline - Ring A = Phenyl

R ₂ NH=4-(4-chlorobenzyl)piperidine or 1-(4-chlorobenzyl)piperazine

Schemes 6 and 7 show the preparation of compounds of formula Ia in which ring A is substituted. Scheme 6 shows the preparation of compounds of formula Ia with a substituted cyclopentyl ring A. Scheme 7 shows the preparation of compounds of formula la with substituted pyrrolidine ring A, wherein substituted pyrrolidine 7b is prepared by treating unsubstituted pyrrolidine 7a (R=H) with an appropriate reagent.

Scheme 6. Synthesis of Substituted Cycloalkyl Ring A

Scheme 7. Synthesis of Substituted Pyrrolidine Ring A

R ₂ N=4-(4-chlorobenzyl)piperidine or l-(4-chlorobenzyl)piperazine

Reagent R′

CH ₂ CHSO ₂ Me (CH ₂ ) ₂ SO ₂ Me

ICH ₂ CONH ₂ CH ₂ CONH ₂

Ac ₂ O Ac

_{ClCOCH2CO2Me ; KOHCOCH2CO2H _}

NaNCO CONH ₂

ClCONMe ₂ CONMe ₂

MsCl Ms

ClSO ₂ NHBOC; HClSO ₂ NH ₂

ClSO ₂ NMe ₂ SO ₂ NMe ₂

Schemes 8 and 9 show the conversion of compounds of formula (Ia) to compounds of formula (I) wherein L and A are different.

Scheme 8. Conversion of primary amines to ureas and amides

Compounds of _formula (I) wherein L is -C(=O)NRa are prepared as shown in Scheme 8 and general procedures C and D below. Compounds of formula (I) where L is -C(=O)- are prepared as shown in Scheme 8 and General Procedures E and F below.

R ₂ N=4-(4-chlorobenzyl)piperidine or 1-(4-chlorobenzyl)piperazine X=C, O, S(O) _n , NR ^x , where n=0-2 and ^Rx are as defined in the definition of heterocyclyl.

Reaction Scheme 8

General Method C: (urea formation with isocyanate)

A 0.1-0.6 M solution of the amine (1 eq) in _CH2Cl2 or _CH2Cl2 and DMF at 0-20 °C was treated with the indicated isocyanate ( _{1.1-2 eq} ) and stirred for 0.5-1.5 h in CH ₂ Partition between Cl ₂ and saturated NaHCO ₃ . The aqueous phase was extracted with _CH2Cl2 , dried and _the extract was concentrated. Crude urea was purified by column chromatography or preparative TLC, or used directly in the next step without purification.

General Method D: (urea formation with isocyanate)

A 0.1-0.6 M solution of the amine (1 eq) in _CH2Cl2 or _CH2Cl2 and DMF at 0-20 °C was treated with the indicated isocyanate ( _{1.1-2 eq} ) and stirred for 0.5-1.5 h in CH ₂ Partition between Cl ₂ and saturated NaHCO ₃ . The aqueous phase was extracted with _CH2Cl2 , dried and _the extract was concentrated. Crude urea was purified by column chromatography or preparative TLC, or used directly in the next step without purification. The free base solution in _CH2Cl2 was _treated with 1N HCl in _Et2O and concentrated to give the hydrochloride salt.

General Procedure E: (Amide Formation Using 1-Hydroxybenzotriazole and 1-(3-Dimethylaminopropyl)-3-Ethylcarbodiimide Hydrochloride)

A solution of 0.1-0.4 M amine (1 eq) and specified carboxylic acid (1.2-1.5 eq) in _CH2Cl2 at 0 °C was treated with 1-hydroxybenzotriazole hydrate ₍ HOBt) (0.2-0.5 eq ) and 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (DEC) (1.3-2 equivalents) were treated continuously, stirred at 0-20°C for 2-72 h, partitioned between _CH2Cl2 and saturated _NaHCO3 , extracted the aqueous phase with _{CH2Cl2 ,} dried and concentrated the extract _. The crude amide was purified by column chromatography and/or preparative TLC.

General Procedure F: (Amide Formation Using 1-Hydroxybenzotriazole and 1-(3-Dimethylaminopropyl)-3-Ethylcarbodiimide Hydrochloride)

A solution of 0.1-0.4 M amine (1 eq) and specified carboxylic acid (1.2-1.5 eq) in _CH2Cl2 at 0 °C was treated with 1-hydroxybenzotriazole hydrate ₍ HOBt) (0.2-0.5 eq ) and 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (DEC) (1.3-2 equivalents) were treated continuously, stirred at 0-20°C for 2-72 h, partitioned between _CH2Cl2 and saturated _NaHCO3 , extracted the aqueous phase with _{CH2Cl2 ,} dried and concentrated the extract _. The crude amide was purified by column chromatography and/or preparative TLC. The free base solution in _CH2Cl2 was _treated with 1N HCl in _Et2O and concentrated to give the hydrochloride salt.

Scheme 9 and the following Methods G-O describe various methods for converting Formula Ia to compounds of Formula I, wherein L varies.

General Procedure G (Parallel Synthesis of Sulfonamides)

A mixture of the desired amine Ia (1 eq), the appropriate sulfonyl chloride (1.5 eq), and AmberliteIRA67 (2 eq) in _CH2Cl2 ( ₂ ml) was rotated overnight. The mixture was treated with PS-trisamine (1.2 equiv) (Argonaut Technologies Inc., San Carlos, CA, USA) and rotated overnight. The solid _was collected by filtration, washed with _CH2Cl2 , _MeOH and _CH2Cl2 . Concentration of the filtrate afforded the product.

General Procedure H (Parallel Synthesis of Amides from Acid Chlorides)

A mixture of the desired amine Ia (1 eq), the appropriate acid chloride (1.5 eq), and Amberlite IRA67 (2 eq) in _CH2Cl2 ( ₂ ml) was rotated overnight. The mixture was treated with PS-trisamine (1.2 equiv) and MP-carbonate (2 equiv) (Argonaut Technologies, San Carlos, CA) and rotated overnight. The solid _was collected by filtration, washed with _CH2Cl2 , _MeOH and _CH2Cl2 . Concentration of the filtrate afforded the product.

General Procedure I (Parallel Synthesis of Amides from Carboxylic Acids)

The desired amine Ia (1 eq), the appropriate carboxylic acid (1.5 eq), and PS-carbodiimide (2 eq) (Argonaut Technologies Inc., San Carlos, CA, USA) were _dissolved in _CH2Cl2 ( 2 ml) was swirled overnight. The mixture was treated with MP-carbonate (2 eq) and rotated overnight. The solid _was collected by filtration, washed with _CH2Cl2 , _MeOH and _CH2Cl2 . Concentration of the filtrate afforded the product.

General Procedure J (Parallel Synthesis of Urea from Isocyanates and Purification by Parallel Chromatography)

A mixture of the desired amine Ia (1 eq) and the appropriate isocyanate (1.2 eq) in _{CH2Cl2 (} 2 ml) was rotated overnight. The _mixture was concentrated to give the crude product, which was purified by parallel chromatography with a step gradient (2.5% MeOH/ _CH2Cl2 , 10% MeOH/ _{CH2Cl2 )} .

General Procedure K (Parallel Synthesis of Urea from Isocyanate and Purification by Catch and Release Scavenger)

A mixture _of the desired amine Ia (1 eq) and the appropriate isocyanate (1.2 eq) in _CH2Cl2 (2 ml) was stirred overnight. The mixture was treated with MP-TsOH and rotated for 3 hours. The solid _was collected by filtration, washed with _CH2Cl2 , _MeOH and _CH2Cl2 . The solid was rotated with 2M _NH3 in MeOH for 2 hours. The solid _was collected by filtration, washed with _CH2Cl2 , _MeOH and _CH2Cl2 . Concentration of the filtrate afforded pure product.

General procedure L (parallel synthesis of urea from aniline using Phoxime resin)

A mixture of the appropriate aniline (3 equiv) and Phoxime resin (1 equiv) in _CH2Cl2 ( _2ml ) was swirled for 3 hours. If the aniline was still not dissolved, triethylamine (3.5 equiv) was added. The mixture was swirled overnight. The solid was _collected by filtration, washed with _CH2Cl2 , MeOH, _CH2Cl2 , _MeOH and _{CH2Cl2 .} A mixture of the solid and the desired amine Ia (1.1 equiv) in _CH2Cl2 ( _0.5ml ) and toluene (1.5ml) was heated to 80°C and shaken overnight, then cooled to room temperature. The solid _was collected by filtration, washed with _CH2Cl2 , _MeOH and _CH2Cl2 . Concentration of the filtrate afforded the product.

General method M (parallel synthesis of urea from aniline using triphosgene)

A mixture of _the appropriate aniline (1.2 equiv), triphosgene (0.4 equiv), and triethylamine (1.4 equiv) in _CH2Cl2 was heated at 35 °C for 1 hour. After cooling to room temperature, the desired amine Ia (1 equiv) was added. The mixture was stirred overnight, washed with water and brine, passed over sodium sulfate, and concentrated to give the crude product, which was purified by parallel chromatography.

General Procedure N (Parallel Synthesis of Thiourea from Thioisocyanate)

A mixture of the desired amine Ia (1 eq) and the appropriate thioisocyanate (1.2 eq) in _CH2Cl2 (2 ml) was stirred _overnight . Treat with MP-TsOH mixture and shake with rotation for 3 hours. The solid _was collected by filtration, washed with _CH2Cl2 , _MeOH and _CH2Cl2 . The solid was vortex shaken with 2M _NH3 in MeOH for 2 h. The solid _was collected by filtration, washed with _CH2Cl2 , _MeOH and _CH2Cl2 . Concentration of the filtrate afforded the purified product.

General Procedure O (Parallel Synthesis of Carbamates)

A mixture of the desired amine Ia (1 eq) and the appropriate succinimide (1.5 eq) in _{CH2Cl2 (} 2 ml) was stirred overnight. If the reaction is not complete, heat at 38°C for 1 hour. The mixture was washed with water and brine, passed through sodium sulfate, and concentrated to give _the crude product which was purified in parallel purification steps (step gradient 5% MeOH/ _CH2Cl2 , 10% MeOH/ _{CH2Cl2 )} .

Unless otherwise stated, all non-aqueous reactions were performed under nitrogen atmosphere and sodium sulfate was used to dry all organic layers. Purification was typically performed by flash chromatography on silica gel (230-400 mesh) or preparative TLC on Uniplate Silica Gel GF PLC plates from Analtech, Inc., Newark, DE. The alumina used was basic and had 6 wt% _H2O (Brockmann III). Melting point tests performed in capillaries are not corrected. IR spectra were measured in KBr. NMR spectra were performed in _CDCl3 unless otherwise stated. ¹ H NMR spectra were recorded on a 300 MHz instrument and ¹³ C NMR spectra were recorded at 75.5 MHz. Mass spectrometry was achieved using electron spray ionization. Analytical reversed-phase HPLC was performed on a Shimadzu system equipped with a diode array spectrometer (range 190-300 nm; Hewlett Packard). The stationary phase was a Zorbax SB-Phenyl Rapid Resolution column (4.6 mm×50 mm; Hewlett Packard), the mobile phase A was 0.1% trifluoroacetic acid, and the mobile phase B was CH ₃ CN. Flow rate 2.5 ml/min, using a linear gradient of 20-55% B over 5 minutes, followed by a linear gradient of 55-20% B over 5 minutes. All parallel assay reactions were performed in locked tubes that had been vortexed overnight. _Wash successively with _{CH2Cl2 ,} MeOH, _CH2Cl2 , MeOH _{, CH2Cl2} and dry under vacuum before use. All products from parallel synthesis reactions were identified by HPLC-MS.

Example

As described in the reaction schemes and examples, the following preparations (1-7) are used to prepare synthetic intermediates that can be used to prepare the compounds of the present invention.

Preparation 1: Preparation of (±)-trans-2-[4-(4-chlorobenzyl)piperidin-1-yl]-cyclohexylamine

Step A: Preparation of (±)-trans-2-[4-(4-chlorobenzyl)piperidin-1-yl]-cyclohexanol

Alkylation of 4-(4-chlorobenzyl) with 7-oxa-bicyclo[4.1.0]heptane (0.25ml, 2.5mmol) in EtOH (0.5ml) at 80°C following method A )-piperidine (see Preparation 7) (52 mg, 0.25 mmol) for 3 days. Chromatography of the crude product with 90:9.5:0.5-80:19: ₁ CH2Cl2: _{MeOH:NH4OH gave} the product as a tan oil (68 mg, 88%) which solidified to a milky solid on standing: mp 100-101.3 ℃; IR 3379, 2929cm ^-1 ; ¹ H NMR δ1.05-1.76(m, 12H), 2.02(dt, J=2.4, 11.6Hz, 1H), 2.06-2.20(m, 2H), 2.49(d, J=7.0Hz, 2H), 2.51-2.64(m, 2H), 2.79(m, 1H), 3.34(m, 1H), 4.05(m, 1H), 7.06(m, 2H), 7.24(m, 2H ); MS m/z 308 (M+H) ⁺ .Anal. (C ₁₈ H ₂₆ ClNO) C, H, N.

Step B: Preparation of (±)-trans-2-[4-(4-chlorobenzyl)piperidin-1-yl]-cyclohexylamine

(±)-trans-2-[4-(4-chlorobenzyl)piperidin-1-yl was treated successively with Et3N (350 μl, 2.53 mmol) and MeSO ₂ Cl (194 μl, 2.53 mmol) at 0 °C A solution of ]-cyclohexanol (390mg, 1.27mmol) in _CH2Cl2 (6ml ₎ , stirred at 0°C for ₂ hours, partitioned between _CH2Cl2 and 10% _NH4OH . The aqueous phase was extracted with _CH2Cl2 , the extract was _washed with brine, dried and concentrated. A solution of the residue in THF (3ml) and 28-30wt% _NH4OH (1.2ml) was stirred at 70°C for 24h, cooled to room temperature and partitioned between EtOAc and 1N NaOH. The aqueous phase was extracted with EtOAc, the extract was washed with brine, dried and concentrated. The residue was chromatographed on alumina with 1:3 EtOAc:MeOH to 100% MeOH, followed by 20:1 Hexane:EtOAc to 100% EtOAc on alumina, followed by 3:1 EtOAc:MeOH to 100% MeOH layer Analysis gave a tan oily product (260 mg, 67%), which solidified after standing: mp 69.1-70.4°C; ¹ H NMR δ 1.03-1.34 (m, 6H), 1.37-1.52 (m, 1H), 1.57 -1.77(m, 5H), 1.92-2.05(m, 3H), 2.48(d, J=7.0Hz, 2H), 2.45-2.64(m, 3H), 2.73(m, 1H), 7.06(m, 2H ), 7.23(m, 2H); MS m/z 307(M+H) ⁺ .

Preparation 2: Preparation of (±)-trans-2-[4-(4-chlorobenzyl)piperidin-1-yl]-cyclopentylamine

Step A: Preparation of (±)-trans-2-[4-(4-chlorobenzyl)piperidin-1-yl]-cyclopentanol

Following method A, 4-(4-chlorobenzyl)-piperidine (17.86 g, 85.05 mmol) and 6-oxa-bicyclo[3.1.0]hexane (50 g, 0.6 mol) were stirred at 95 °C in Solution in EtOH (170ml) for 40h, cooled to room temperature and concentrated. The residue was crystallized in hot _CH2Cl2 ( _80ml ), the crystallized mixture was concentrated to half volume, kept at 0°C overnight, filtered, and the precipitate rinsed with cold hexane to give the product as a tan solid (18.2g, 73%). The mother liquor was concentrated to half volume, diluted with _CH2Cl2 , kept at -10 °C for 1 h, and the precipitate _was rinsed with cold _{CH2Cl2 and} hexanes to give additional product as a tan solid (1.8 g, 7%) : mp 104.1-105.5°C; IR 3436, 2928cm ^-1 ; ¹ HNMR δ1.19-1.75(m, 8H), 1.81-1.99(m, 4H), 2.06(dt, J=2.5, 11.7Hz, 1H), 2.47(m, 1H), 2.50(d, J=7.0Hz, 2H), 2.90(m, 1H), 3.07(m, 1H), 4.10(m, 1H), 7.06(m, 2H), 7.23(m , 2H); 13C NMR 8 21.63, 27.35, 32.01, 32.15, 34.31, 37.87, 42.47, 50.47, 52.97, 75.15, 75.22, 128.27, 130.43, 131.55, 139.04; MSm/z 294(M+H) ⁺ .Anal. (C ₁₇ H ₂₄ ClNO.0.1H ₂ O)C, H, N.

Step B: Preparation of (±)-trans-2-[4-(4-chlorobenzyl)piperidin-1-yl]-cyclopentylamine

Following general procedure B, (±)-trans-2-[4-(4-chlorobenzyl) was treated successively with _Et3N (190 μl, 1.4 mmol) and _MeSO2Cl (110 μl, 1.4 mmol) at 0 °C yl)piperidin-1-yl]-cyclopentanol (205 mg, 0.697 mmol) in CH ₂ Cl ₂ (2.8 ml), stirred at 0° C. for 1 h, in CH ₂ Cl ₂ and 10% NH ₄ Allocation among OH. The aqueous phase was extracted with _CH2Cl2 , the extract was washed with brine, dried and concentrated to give 220 mg of _an oil. A solution of the residue (110 mg) in dioxane (2 ml) and 28-30 wt% _NH4OH (0.8 ml) was stirred at 70-80 °C overnight, cooled to room temperature, and concentrated. The residue was partitioned between EtOAc and 1N NaOH, the aqueous phase was extracted with EtOAc, the extract was washed with brine, dried and concentrated. Chromatography of the residue on alumina with 10 _: 1 hexanes:EtOAc to 100% EtOAc followed by 95:4.75:0.25-60:38 _:2 CH2Cl2:MeOH _:NH4OH gave the product as an oil (87 mg, 85%): ¹ H NMR δ1.18-1.71 (m, 9H), 1.76-2.00 (m, 3H), 2.07 (dt, J=2.4, 11.5Hz, 1H), 2.31 (m, 1H), 2.50 (d , J=6.9Hz, 2H), 2.86-2.99(m, 2H), 3.19(m, 1H), 7.06(m, 2H), 7.23(m, 2H); MS m/z293.2(M+H) ⁺ .

Preparation 3: Preparation of (±)-cis-2-[4-(4-chlorobenzyl)piperidin-1-yl]-cyclopentylamine

Step A: Preparation of (±)-trans-4-nitro-benzenesulfonic acid 2-azido-cyclopentyl ester

(±)-trans-2-azido-cyclopentanol (1.27 g , 10.0 mmol) (Zhang, Z.da; Scheffold, R. Helv. Chim. Acta 1993, 76, 2602) in CH ₂ Cl ₂ (14 ml) was slowly warmed to room temperature. The residue was stirred for 4 days, during which time additional pyridine (0.9ml, 11mmol) and 4-nitro-benzenesulfonic acid (2.2g, 10mmol) were added, partitioned between _CH2Cl2 and _1N HCl. The aqueous phase was extracted with _CH2Cl2 , the extract was washed with saturated _NaHCO3 , _dried and concentrated. The residue was chromatographed with 10:1-4:1 hexanes:EtOAc to give the product as a yellow oil (2.63 g, 84%): ¹ H NMR δ 1.61-1.90 (m, 4H), 2.00-2.16 (m, 2H), 3.96(m, 1H), 4.72(m, 1H), 8.14(m, 2H), 8.43(m, 2H).

Step B: Preparation of (±)-cis-1-(2-azido-cyclopentyl)-4-(4-chlorobenzyl)-piperidine

(±)-trans-4-nitro-benzenesulfonic acid 2-azido-cyclopentyl ester (630mg, 2.0mmol), 4-(4-chlorobenzyl)-piperidine (420mg, 2.0mmol ) and _Et3N (280 μl, 2.0 mmol) in _CH3CN (4 ml) was stirred at room temperature for 10 days, at 65 °C for 2 days, cooled to room temperature, and concentrated. The residue was partitioned between _CH2Cl2 and 1N HCl, the aqueous phase was extracted with _CH2Cl2 , dried _and the extract was concentrated _. Chromatography _of the residue with 20:1-1:1 hexane:EtOAc followed by 100% _CH2Cl2 to 95:4.75: _{0.25 CH2Cl2} :MeOH: _NH4OH afforded the product as a tan oil (145 mg , 22%): ¹ H NMR δ1.32-1.90 (m, 13H), 2.33 (m, 1H), 2.49 (d, J=6.4Hz, 2H), 2.96 (m, 1H), 3.06 (m, 1H ), 4.04(t, J=4.0Hz, 1H), 7.06(m, 2H), 7.23(m, 2H); MS m/z 319.2(MH) ^- .

Step C: Preparation of (±)-cis-2-[4-(4-chlorobenzyl)piperidin-1-yl]-cyclopentylamine

₍ ±)-cis-1-(2-azido-cyclopentyl)-4-(4-chlorobenzyl)- A solution of piperidine (210 mg, 0.65 mmol) in THF (2.5 ml) was refluxed for 3.5 hours, cooled to room temperature, and concentrated. Chromatography with 90:9.5:0.5-75:23.75:1.25 _CH2Cl2 : _MeOH : _NH4OH gave the product as a colorless oil (183 mg, 95%) which solidified to a milky solid on standing: mp 69.6-71.3 ℃; ¹ H NMRδ1.20-1.35 (m, 2H), 1.43-1.93 (m, 11H), 2.17 (m, 1H), 2.49 (d, J=6.9Hz, 2H), 2.89-3.02 (m, 2H ), 3.34(t, J=4.4Hz, 1H), 7.06(m, 2H), 7.23(m, 2H); 128.63, 130.80, 131.88, 139.58; MS m/z 293.2(M+H) ⁺ .

Preparation 4: Preparation of (±)-cis-2-[4-(4-chlorobenzyl)piperidin-1-yl]-cyclohexylamine

Step A: Preparation of (±)-trans-4-nitro-benzenesulfonic acid 2-azido-cyclohexyl ester

(±)-trans-2-azido-cyclohexan-1-ol (11.3 g, 80.0 mmol) (Zhang, Z. da; Scheffold, R. Helv. Chim. Acta 1993, 76, 2602) in CH ₂ Cl ₂ (110 ml) was slowly warmed to room temperature. The residue was stirred for 4 days, partitioned between _CH2Cl2 and 1N HCl _. The aqueous phase was extracted with _CH2Cl2 , the extract was washed with saturated _NaHCO3 , _dried and concentrated. Chromatography of the residue with 10:1-1:1 hexanes:EtOAc gave the product as a milky solid (19 g, 72%): ¹ H NMR δ 1.19-1.39 (m, 3H), 1.53-1.82 (m, 3H), 2.00 -2.10(m, 1H), 2.26(m, 1H), 3.36(m, 1H), 4.35(ddd, J=4.7, 9.2, 10.8Hz, 1H), 8.17(m, 2H), 8.41(m, 2H ).

Step B: Preparation of (±)-cis-1-(2-azido-cyclohexyl)-4-(4-chloro-benzyl)-piperidine

(±)-trans-4-nitro-benzenesulfonic acid 2-azido-cyclohexyl ester (1.77g, 5.41mmol), 4-(4-chlorobenzyl)-piperidine (1.14g, 5.43 mmol), and a cloudy solution of _Et3N (0.75ml, 5.4mmol) in _CH3CN (11.2ml) was stirred at room temperature for 17 hours, at 65°C for 31 hours, and at 80°C for 5 days and cooled to room temperature , and concentrate. The residue was partitioned between _CH2Cl2 and 1N NaOH, the aqueous phase was extracted with _CH2Cl2 , dried _and the extract was concentrated _. The residue was chromatographed with 98:1.9:0.1-95 _:4.75 :0.25 _CH2Cl2 :MeOH _: NH4OH to 100% MeOH, followed by 10:1 Hex:EtOAc to 100% EtOAc, then 95:5 EtOAc : MeOH chromatography afforded sequentially eluting starting (±)-trans-4-nitro-benzenesulfonic acid 2-azidocyclohexyl ester (1.2 g, 68%), desired product (155 mg, 9 %), and starting 4-(4-chlorobenzyl)-piperidine (810 mg, 71%). Product: ¹ HNMRδ1.19-1.81(m, 12H), 1.92-2.08(m, 3H), 2.22(m, 1H), 2.48(d, J=7.0Hz, 2H), 3.02(m, 2H), 4.05 (m, 1H), 7.06(m, 2H), 7.23(m, 2H); MS m/z333.2(M+H) ⁺ .

Step C: Preparation of (±)-cis-2-[4-(4-chlorobenzyl)piperidin-1-yl]-cyclohexylamine

(±)-cis- _1- (2-azido-cyclohexyl)-4-(4-chlorobenzyl)-piperene was treated sequentially with PPh (364 mg, 1.39 mmol) and water (141 μl, 5.56 mmol) A solution of pyridine (155mg, 0.463mmol) in THF (1.8ml) was refluxed for 3 hours, cooled to room temperature, and concentrated. Chromatography with 95:4.75:0.25-75:23.75:1.25 CH ₂ Cl ₂ :MeOH:NH ₄ OH gave the product as a milky solid (121 mg, 85%): ¹ H NMR δ 1.14-1.93 (m, 15H), 1.96 ( dt, J=11.8, 3.5Hz, 1H), 2.48(d, J=7.0Hz, 2H), 3.03-3.13(m, 2H), 3.30(m, 1H), 7.06(m, 2H), 7.23(m , 2H); MS m/z307.2(M+H) ⁺ .

Preparation 5: Preparation of (±)-trans-2-[4-(4-chlorobenzyl)piperidin-1-yl]-cyclobutylamine

Step A: Preparation of (±)-2-[4-(4-chlorobenzyl)piperidin-1-yl]-cyclobutanone

1,2-Bis( Trimethylsilyloxy)cyclobutene (5.0 g, 22 mmol) and warmed to room temperature. The reaction was stirred over 5 hours during which time additional 1,2-bis(trimethylsilyloxy)cyclobutene (0.99 g, 4.3 mmol) was added and concentrated. Chromatography of the residue with 95:4.75:0.25 CH ₂ Cl ₂ :MeOH:NH ₄ OH afforded the product as a yellow oil (4.8 g, 80%): ¹ H NMR δ 1.20-1.35 (m, 2H), 1.43-1.64 (m , 3H), 1.93-2.18(m, 4H), 2.49(d, J=6.9Hz, 2H), 2.64-2.91(m, 3H), 3.14(m, 1H), 3.90(m, 1H), 7.05( m, 2H), 7.23(m, 2H); MS m/z 278.1(M+H) ⁺ .

Step B: Preparation of (±)-2-[4-(4-chlorobenzyl)piperidin-1-yl]-cyclobutanone O-methyl-oxime

(±)-2-[4-(4-Chlorobenzyl)piperidin-1-yl]-cyclobutanone (1.74 g, 6.26 mmol) and MeONH ₂ .HCl (2.63 g, 31.3 mmol) in MeOH (20ml) was stirred at 65°C for 3 hours, cooled to room temperature, and concentrated. The _residue was partitioned between _CH2Cl2 and saturated _NaHCO3 , the aqueous phase was extracted with _CH2Cl2 , the extracts _were dried and concentrated. Chromatography of the residue with 95:4.75:0.25 CH ₂ Cl ₂ :MeOH:NH ₄ OH gave the product (1.5 g, 78%) as a stereoisomer-predominant brown oil: ¹ H NMR δ 1.05-1.65 (m , 4.5H), 1.92-2.11(m, 4H), 2.45-2.65(m, 3H), 2.73-2.96(m, 2H), 3.22(m, 1H), 3.73(m, 1H), 3.82(m, 3H), 4.57(m, 0.5H), 7.06(m, 2H), 7.23(m, 2H); MS m/z 307.1(M+H) ⁺ .

Step C: Preparation of (±)-trans-2-[4-(4-chlorobenzyl)piperidin-1-yl]-cyclobutylamine

Under an atmosphere of argon, a mixture of NaBH ₄ (604 mg, 16.0 mmol) in THF (13 ml) was treated dropwise with trifluoroacetic acid (1.23 ml, 16.0 mmol), stirred for 5 minutes, and washed dropwise with (±)-2-[ 4-(4-Chlorobenzyl)piperidin-1-yl]-cyclobutanone O-methyl-oxime (985mg, 3.21mmol) was treated with a solution in THF (35ml) and stirred at room temperature for 5 hours . The mixture was carefully treated with 6N HCl (1.5ml) until the pH was about 2, stirred for 10 min, basified with 8N NaOH until the pH was about 10, and partitioned between EtOAc and 1N NaOH. The aqueous phase was extracted with EtOAc, the extracts were washed with brine, _dried ( _Na2SO4 ) and concentrated. A solution of the residue in MeOH (30ml) and 1N HCl (3ml) was stirred at 50°C for 1 hour, at 75°C for 5 hours, cooled to room temperature, and concentrated. _The residue was partitioned between _CH2Cl2 and 1N NaOH, the aqueous phase was extracted with _{CH2Cl2 ,} dried and the extract was concentrated. Chromatography of the residue on alumina with 10:1 hexanes:EtOAc to 100% EtOAc followed by 98:1.9:0.1-90 _:9.5 : _{0.5 CH2Cl2} :MeOH _:NH4OH gave the product as a yellow oil ( ¹ H NMR 80% purity), which was used without further purification: ¹ H NMR δ 1.19-1.90 (m, 9H), 2.11 (m, 1H), 2.28 (m, 1H), 2.44-2.59 (m, 3H), 2.80(m, 1H), 3.05(m, 1H), 3.22(m, 1H), 7.06(m, 2H), 7.23(m, 2H); MS m/z 279.2(M+H) ⁺ .

Preparation 6: Preparation of (±)-cis-2-[4-(4-chlorobenzyl)piperidin-1-yl]-cyclobutylamine

(±)-2-[4-(4-Chlorobenzyl)piperidin-1-yl]-cyclobutanone O-methyl-oxime (438mg, 1.43mmol) in THF (13ml) under argon atmosphere A solution of <RTI ID=0.0>(R)</RTI> was treated dropwise with 1M _BH3.THF complex in THF (8.6ml, 8.6mmol) and stirred at room temperature for 3 hours and at 75°C for 20 hours. Carefully treat with 6N HCl (1 ml) until the pH is about 2. The THF was evaporated and a solution of the residue in EtOH (9ml) and 6N HCl (1ml) was stirred at 75°C for 1 hour. It was then cooled to room temperature, basified with 8N NaOH (4ml) until the pH was about 10, diluted with water (5ml) until the resulting white precipitate dissolved, and concentrated. The residue was partitioned between _CH2Cl2 and 1N NaOH, the aqueous phase was extracted with _{CH2Cl2 ,} dried and the extract was concentrated _. Chromatography of the residue with 90:9.5:0.5-60:38:2 CH ₂ Cl ₂ :MeOH:NH ₄ OH afforded 70 mg of the desired product ( ¹ HNMR 80% purity) sequentially eluting as a colorless oil, which was used without After further purification, 48 mg (12%) of pure desired product as a colorless oil, and 125 mg of desired product, stereoisomeric (±)-trans-2-[4-(4-chlorobenzyl)piperidine Mixture of -1-yl]-cyclobutylamine and unidentified impurities. Product: ¹ H NMR δ1.19-1.70(m, 8H), 1.89-2.05(m, 3H), 2.50(d, J=6.9Hz, 2H), 2.56(m, 1H), 2.78(m, 2H) , 3.44 (m, 1H), 7.06 (m, 2H), 7.23 (m, 2H); 13C NMR 824.39, 25.56, 31.63, 31.76, 38.01, 42.61, 49.17, 49.63, 51.74, 62.51, 128.25, 130.42, 131.50, 139.16; MS m/z 279.2(M+1) ⁺ .

Preparation 7: Preparation of 4-(4-chlorobenzyl)-piperidine

Step A: Preparation of tert-butyl 4-(4-chloro-benzylidene)-piperidine-1-carboxylate

The phosphonium salt (10 g) was taken in THF and placed on an ice bath. KHMDS (42ml) was added slowly, the ice bath was removed, and the reaction was stirred at room temperature for 45 minutes. Then the reaction solution was cooled to -78°C, and ketone (4.2 g) was added slowly. The reaction was stirred for 30 minutes, the cooling bath was removed, and stirred overnight at room temperature. The reaction solution was poured into saturated _NH4Cl (100ml) solution, the layers were separated, the aqueous layer was washed twice with EtOAc, the organic layers were combined, dried (MgSO4), and concentrated to about 40ml. The solution was diluted with hexanes and filtered to remove the major _Ph3PO . The crude product was chromatographed with 20:1-10:1 hexanes:EtOAc to give the product as a colorless oil (4.7g).

Step B: Preparation of tert-butyl 4-(4-chloro-benzyl)-piperidine-1-carboxylate

The protected piperidine (10 g) was dissolved in EtOAc (100 ml), _PtO2 was added, and the mixture was stirred under hydrogen atmosphere for 3 hours. The mixture was filtered through celite and concentrated. The crude product was taken up in hot hexanes, filtered, and allowed to crystallize. Recrystallization of the product from hot hexanes gave pure product (8.0 g). Additional product was isolated from the mother liquor.

Step C: Preparation of 4-(4-chlorobenzyl)piperidine

Methanol (400ml) was placed in an ice bath and AcCl (60ml) was added. After the addition was complete, the solution was stirred at room temperature for 1 hour. The protected piperidine (62.8 g) was added and the solution was stirred overnight at room temperature. The reaction solution was concentrated to about 70ml (when the product started to precipitate out), diluted with diethyl ether (500ml) and the product was collected by filtration (44.9g). An additional 3.1 g of product was collected from the mother liquor.

Example 1: Preparation of the following compound cyclohexanecarboxylic acid {(1R,2R)-2-[4-(4-chloro-benzyl)-piperidine using the appropriate amine 1a and carboxylic acid using general procedure E -1-yl]-cyclopentyl}-amide

Example 2: Application of General Procedure K using the appropriate amine 1a and the isocyanate ^R4N =C=O to prepare the following compounds

1-allyl-3-{(1R,2R)-2-[4-(4-chloro-benzyl)-piperidin-1-yl]-cyclopentyl}-urea;

1-{(1R,2R)-2-[4-(4-Chloro-benzyl)-piperidin-1-yl]-cyclopentyl}-3-isopropyl-urea;

1-Butyl-3-{(1R,2R)-2-[4-(4-chloro-benzyl)-piperidin-1-yl]-cyclopentyl}-urea; and

3-(3-{(1R,2R)-2-[4-(4-Chloro-benzyl)-piperidin-1-yl]-cyclopentyl}-ureido)-propionic acid ethyl ester;

Example 3: Application of General Procedure J using the appropriate amine 1a and the isocyanate ^R4N =C=O to prepare the following compounds

{(1R,2R)-2-[4-(4-Chloro-benzyl)-piperidin-1-yl]-cyclopentyl}-3-cyclohexyl-urea.

Embodiment 4 preparation embodiment

The following are representative pharmaceutical formulations containing compounds of formula (I).

Tablet preparation

The following ingredients are mixed thoroughly and compressed into single scored tablets.

                                           

The compound of the present invention 400

                                                         

    Croscarmellose Sodium                    

Lactose 120

                                                       

Capsule preparation

Mix the following ingredients well and fill into a hard shell gelatin capsule.

Amount per capsule, mg

The compound of the present invention 200

Lactose, spray-dried 148

Magnesium Stearate 2

Suspension preparation

The following ingredients are mixed to form a suspension for oral administration.

Ingredient Quantity

The compound of the present invention 1.0g

                                             

Sodium Chloride 2.0g

Methylparaben 0.15g

    Propyl paraben           0.05g

                                                                                                            

Sorbitol (70% solution) 12.85g

Veegum K (Vanderbilt Company) 1.0g

                                                                       

                                                                                           

Distilled water Make up to 100ml

Injection preparation

The following ingredients are mixed to form an injection preparation.

Ingredient Quantity

The compound of the present invention 0.2g

Sodium acetate buffer solution, 0.4M 2.0ml

  HCl(1N) or NaOH(1N) Supplement to proper pH

Water (distilled, sterile) Make up to 20ml

Liposome formulation

The following ingredients are mixed to form a liposome formulation.

Ingredient Quantity

The compound of the present invention 10mg

L-.α.-Phosphatidylcholine 150mg

                                     

Samples were freeze-dried and lyophilized overnight. Samples were reconstituted with 1 ml of 0.9% saline solution. The particle size of liposomes can be reduced by sonication.

Example 5: In vitro CCR-3 receptor binding detection

The CCR-3 antagonistic activity of the compounds of the present invention can be determined by their ability to inhibit the binding of ¹²⁵ Ieotaxin to CCR-3 L1.2 transfected cells (see Ponath, PD et al., J. Exp. Med., Vol. 183, 2437- 2448, (1996)).

Assays were performed in Costar 96-well polypropylene round bottom plates. Test compounds were dissolved in DMSO, and then diluted with binding buffer (50 mM HEPES, 1 mM CaCl.sub.2, 5 mM MgCl ₂ , 0.5% bovine serum albumin (BSA), 0.02% sodium azide, pH 7.24) so that The final concentration of DMSO was 2%. 25 μl of test solution or buffer containing only DMSO (control sample) was added to each well, followed by 25 μl of ¹²⁵ I-eotaxin (100 pmol) (NEX314, New England Nuclear, Boston, Mass.) and 25 μl of binding buffer 1.5 x 10 ⁵ CCR-3 L1.2 transfected cells. The final reaction volume was 75 μl.

After incubating the reaction mixture for 1 hour at room temperature, the reaction was terminated by filtering the reaction mixture through polyethylenimine-treated Packard Unifilter GF/C filter plates (Packard, Chicago, 111.). Filters were washed 4 times with ice-cold wash buffer (pH 7.2) containing 10 mm HEPES and 0.5 M NaCl, and dried at 65° C. for about 10 minutes. 25 [mu]l/well Microscint-20( ^R ) scintillation fluid (Packard) was added and the radioactivity retained on the filters was determined by using a Packard TopCount( ^R) .

Compounds of the invention are active in this test.

Example 6: Inhibition of Eotaxin-mediated chemotaxis of CCR-3 L1.2 transfected cells-in vitro test

The CCR-3 antagonistic activity of the compounds of the present invention can be determined by measuring the inhibition of eotaxin-mediated chemotaxis of CCR-3 L1.2 transfected cells using a slightly modified method, in Ponath, PD et al., J. Clin. This method is described in Invest. 97:604-612 (1996). Assays were performed in 24-well chemotaxis dishes (Collaborative Biomedical Products). CCR-3 L1.2 transfected cells were grown in culture medium containing RPMI 1640, 10% Hyclone ^(R) fetal bovine serum, 55 mM 2-mercaptoethanol and Geneticin 418 (0.8 mg/ml). 18-24 h before the assay, treat transfected cells with n-butyric acid at a final concentration of 5 mM/1 x ¹⁰⁶ cells/ml, detach the cells and resuspend in assay medium at a level of 1 x ¹⁰⁷ cells/ml , the assay medium contained equal parts of RPMI 1640 and medium 199 (M199) with 0.5% bovine serum albumin.

Add 1 mg/ml of human eotaxin suspended in phosphate buffer to the bottom chamber to make a final concentration of 100 nM. Transwell culture inserts (Costar Corporation, Cambridge, Mass.) with a pore size of 3 microns were inserted into each well, and L1.2 cells (1×10 ⁶ ) were added to the apical chamber in a final volume of 100 μl. Test compounds in DMSO were added to both the top and bottom chambers such that the final DMSO volume was 0.5%. Assays are performed relative to two sets of controls. Positive controls contained cells in the apical chamber, no test compound, and eotaxin in the lower chamber. Negative controls contained cells without test compound in the apical chamber and neither eotaxin nor test compound in the lower chamber. Plates were incubated at 37°C. After 4 hours, the insert was removed from the chamber and the cells that had migrated to the bottom chamber were counted by pipetting 500 μl of the cell suspension from the lower chamber into a 1.2 ml Cluster tube (Costar) and counting on FACS for 30 seconds to count them.

Example 7: Inhibition of Eotaxin-mediated Chemotaxis of Human Eosinophils - In Vitro Test

The ability of compounds of the present invention to inhibit eotaxin-mediated chemotaxis of human eosinophils can be assessed using a slightly modified method described in Carr, MW et al., Proc.Natl.Acad.Sci.USA, 91:3652- 3656 (1994) as described. Experiments were performed using 24-well chemotaxis plates (Costar Corporation, Cambridge, Mass.). Eosinophils were isolated from blood using the method described in PCT Application, Publication No. WO 96/22371. The endothelial cells used were the endothelial cell line ECV 304 obtained from the European Animal Cell Culture Collection (Porton Down, Salisbury, UK). Endothelial cells were cultured on Biocoat.RTM.Transwell tissue culture inserts (Costar Corporation, Cambridge MA) with a diameter of 6.5 mm and a pore size of 3.0 μM. The medium for ECV 304 cells consisted of M199, 10% fetal bovine serum, L-glutamine and antibiotics. Assay medium consisted of equal parts RPMI 1640 and M199 and 0.5% bovine serum albumin. 24 hours prior to the assay, 2 x ¹⁰⁵ ECV 304 were plated on each insert of a 24-well chemotaxis dish and incubated at 37°C. 20 nM eotaxin diluted in assay medium was added to the bottom chamber for a final volume of 600 μl. Insert endothelial cell-covered tissue culture inserts into each well. Add ¹⁰⁶ eosinophils suspended in 100 μl assay buffer to the apical chamber. Test compounds were dissolved in DMSO and added to both the top and bottom chambers such that the final DMSO volume in each well was 0.5%. Assays were performed relative to two sets of controls. Positive controls contain cells in the apical chamber and eotaxin in the lower chamber. Negative controls contained cells in the top chamber and only assay buffer in the bottom chamber. Chemotaxis dishes were incubated at 37°C for 1-1.5 hours in 5% _CO2 /95% air.

Cells that migrated to the bottom lumen were counted by flow cytometry. 500 μl of the cell suspension was removed from the lower chamber, transferred to a tube, and relative cell counts were obtained by taking the results over a series of 30 second time periods.

Example 8: Inhibition of eosinophil influx by CCR-3 antagonists in the lungs of ovalbumin-sensitized Balb/c mice - in vivo

The ability of compounds of the invention to inhibit infiltration of leukocytes into the lung can be measured by eosinophils in bronchoalveolar lavage (BAL) fluid of ovalbumin (OA)-sensitized balb/c mice following challenge with aerosol antigen The inhibition of aggregation was determined. Briefly, male balb/c mice weighing 20-25 g were sensitized intraperitoneally on days 1 and 14 with OA (10 μg in 0.2 ml aluminum hydroxide solution). After 1 week, the mice were divided into 10 groups. Test compounds or vehicle alone (control group) or anti-eotaxin antibody (positive control) were administered intraperitoneally, subcutaneously or orally. One hour later, the mice were placed in a Plexiglas box and exposed for 20 minutes to OA aerosol generated by a PARISTAR.TM. nebulizer (PARI, Richmond, Va.). Unsensitized or challenged mice were included as negative controls. After 24 or 72 hours, the mice were anesthetized (urethane, approximately 1 g/kg, i.p.), cannulated catheters (PE 60 tubular material) were inserted, and the lungs were lavaged 4 times with 0.3 ml PBS. BAL fluid was transferred into plastic tubes and kept on ice. Total leukocytes in 20 μl aliquots of BAL fluid were measured with a Coulter Counter.TM. Differential leukocyte counts were performed by light microscopy on Cytospin.TM. preparations that had been stained with a modified Wright leukocyte stain (DiffQuick.TM.) using general morphology standards.

The foregoing invention has been described in detail by way of illustration and example for purposes of clarity and understanding. It will be apparent that changes and improvements may be made by a person skilled in the art within the scope of the appended claims. Accordingly, it should be understood that the foregoing description is intended to be illustrative and not restrictive. The scope of the invention should, therefore, be defined not with reference to the foregoing description, but should be determined with reference to the following appended claims, along with the full scope of equivalents to which such claims are entitled.

All patents, patent applications and publications cited herein are incorporated by reference in their entirety to the same extent as if each individual patent, patent application or publication was individually indicated.

Claims (15)

1. Compounds of formula (I): in: R ¹ is (C ₁ -C ₂ ) alkylene; R ² is a benzene ring substituted by one or two substituents selected from C _1-6 alkyl, C _1-6 alkoxy, halogenated C _1-6 alkyl or halogen; R ³ is hydrogen, C _1-6 alkyl, acyl, aryl, or aryl C _1-6 alkyl; Ring A is cyclopentyl; D is N or CR ^b ; L is -C(=O)-, -C(=S)-, -C(=O)N(R ^a )-, -C(=S)N(R ^a )-, -SO ₂ N(R ^a )-, -C(=O)O-, -C(=S)O-; R ⁴ is C _1-6 alkyl, C _3-7 cycloalkyl, C _2-6 alkenyl, C _2-6 alkynyl, heteroalkyl or acyl C _1-6 alkyl; R ^is hydrogen, C _1-6 alkyl, acyl, aryl, aryl C _1-6 alkyl, C _1-6 alkoxycarbonyl, or benzyloxycarbonyl; and R ^b is hydrogen or C _1-6 alkyl; Provided that N-(4-((4-(2-acetylamino-4-fluorophenyl)piperazin-1-yl)methyl)phenylmethyl)acetamide, N-(3,4- Dimethoxyphenylethyl)-N'-(2-acetylaminophenyl)-piperazine, N-[2-(4-benzylpiperazin-1-yl)phenyl]methanesulfonamide and N-Benzyl-N'-(2-acetylaminophenyl)-piperazine, and esters or carbamates, individual stereoisomers, racemic and non-racemic mixtures of stereoisomers, and pharmaceutically acceptable salts of hydroxyl functional groups in compounds of formula (I), in: "Acyl" means -C(O)R group, wherein R is hydrogen, C _1-6 alkyl, C _3-7 cycloalkyl, C _3-7 cycloalkylC _1-6 alkyl, phenyl or Phenyl C _1-6 alkyl; "Acyl C _1-6 alkyl" refers to -C _1-6 alkylene-C (O) R group, wherein R is hydrogen, C _1-6 alkyl, halogenated C _1-6 alkyl, C _{3 -7} cycloalkyl, C _3-7 cycloalkyl-C _1-6 alkyl, optionally substituted phenyl, benzyl, hydroxyl, C _1-6 alkoxy, amino, mono C _1-6 alkane Baseamino or two C _1-6 alkylamino; "Optionally substituted phenyl" refers to phenyl optionally substituted by one or more substituents selected from C _1-6 alkyl, halogenated C _1-6 alkyl, hydroxy C _1-6 Alkyl, heteroalkyl, acyl, amido, amino, C _1-6 alkylamino, diC _1-6 alkylamino, C _1-6 alkylsulfinyl, C _1-6 alkoxy, halogen C _1-6 alkoxy, C _1-6 alkoxycarbonyl, carbamoyl, hydroxyl, halogen, methylenedioxy or ethylenedioxy; "Aryl" refers to a monocyclic or bicyclic aromatic hydrocarbon group of 6-10 ring atoms, which is optionally substituted by one or more substituents selected from C _1-6 alkyl, halogenated C _{1- 6} alkyl, hydroxy C _1-6 alkyl, heteroalkyl, acyl, amido, amino, C _1-6 alkylamino, di-C _1-6 alkylamino, C _1-6 alkylsulfinyl, C _1-6 alkoxy, halogenated C _1-6 alkoxy, C _1-6 alkoxycarbonyl, carbamoyl, hydroxy, halogen, methylenedioxy or ethylenedioxy; "Heteroalkyl" means an alkyl group in which one, two or three hydrogen atoms are replaced by substituents independently selected from -OR ^a and -NR ^b R ^c , provided that the point of attachment of the heteroalkyl is through carbon atom, wherein R ^a is hydrogen, acyl, C _1-6 alkyl, C _3-7 cycloalkyl or C _3-7 cycloalkyl C _1-6 alkyl; R ^b and R ^c are hydrogen independently of each other, Acyl, C _1-6 alkyl, C _3-7 cycloalkyl or C _3-7 cycloalkylC _1-6 alkyl. 2. A compound according to claim 1 which is a compound of formula (II): wherein R ¹ -R ⁴ , A, D and L are as defined in claim 1. 3. A compound according to claim 1 which is a compound of formula (III):

wherein R ¹ -R ⁴ , A, D and L are as defined in claim 1. 4. A compound according to any one of claims 1-3, wherein R ¹ is methylene. 5. Compounds according to any one of claims 1-3, wherein ^R2 is 4-chlorophenyl or 3,4-dichlorophenyl. 6. A compound according to any one of claims 1-3, wherein ^R3 is hydrogen. 7. The compound according to any one of claims 1-3, wherein L is -C(=O)-, -C(=O)N(R ^a )-, -C(=S)N(R ^a ) -, or -C(=O)O-. 8. A compound according to claim 1 which is a compound of formula (IV):

wherein R ³ -R ⁴ , A, D and L are as defined in claim 1. 9. A compound according to any one of claims 1-8, wherein R ⁴ is cyclohexyl, allyl, isopropyl, n-butyl, or 2-(ethoxycarbonyl)ethyl. 10. The compound according to claim 1, which is: Cyclohexanecarboxylic acid {(1R,2R)-2-[4-(4-chloro-benzyl)-piperidin-1-yl]-cyclopentyl}-amide; {(1R,2R)-2-[4-(4-Chloro-benzyl)-piperidin-1-yl]-cyclopentyl}-3-cyclohexyl-urea; 1-allyl-3-{(1R,2R)-2-[4-(4-chloro-benzyl)-piperidin-1-yl]-cyclopentyl}-urea; 1-{(1R,2R)-2-[4-(4-Chloro-benzyl)-piperidin-1-yl]-cyclopentyl}-3-isopropyl-urea; 1-Butyl-3-{(1R,2R)-2-[4-(4-chloro-benzyl)-piperidin-1-yl]-cyclopentyl}-urea; 3-(3-{(1R,2R)-2-[4-(4-Chloro-benzyl)-piperidin-1-yl]-cyclopentyl}-ureido)-propionic acid ethyl ester; or its salt. 11. A process for the preparation of compounds of formula (I) according to claim 1, wherein L is -C(=O)NR ^a -, R ^a is hydrogen, Including the compound of formula (Ia)

Reaction with isocyanates of formula: R ⁴ -N=C=O, wherein R ¹ -R ⁴ , A and D are as defined in claim 1 . 12. A process for the preparation of compounds of formula (I) according to claim 1, wherein L is -C(=O)-, Including the compound of formula (Ia)

Reaction with a compound of formula: R ⁴ -C(=O)OH, wherein R ¹ -R ⁴ , A and D are as defined in claim 1 . 13. A composition comprising a compound or a salt thereof according to any one of claims 1-10 and an excipient. 14. Use of a compound of formula (I) or a salt thereof according to any one of claims 1-10 in the preparation of a medicament for treating a disease treatable by a CCR-3 receptor antagonist. 15. Use according to claim 14, wherein said disease is asthma.